Inhibition of protein kinase C-dependent protein phosphorylation correlates with increased polarity and locomotion in Walker 256 carcinosarcoma cells

Niggli, Verena; Zimmermann, Arthur; Keller, H. U.

doi:10.1002/(sici)1097-0215(19960208)65:4<473::aid-ijc14>3.0.co;2-b

Cited by 11 publications

(22 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Protein kinase C (PKC) isoforms have been shown to regulate ezrin and uropods (Ng et al, 2001;Niggli et al, 1996;Ren et al, 2009;Rossy et al, 2007;Simons et al, 1998;Wald et al, 2008). The PKC family comprises the conventional types, activated by diacylglycerol (DAG) in a Ca 2+ -dependent manner, novel types, activated by DAG but Ca 2+ -independent, and the atypical types, which do not require DAG or Ca 2+ for activation (Mellor and Parker, 1998).…”

Section: Rock and Novel Pkc Isoforms Regulate The Erulsmentioning

confidence: 99%

“…The PKC family comprises the conventional types, activated by diacylglycerol (DAG) in a Ca 2+ -dependent manner, novel types, activated by DAG but Ca 2+ -independent, and the atypical types, which do not require DAG or Ca 2+ for activation (Mellor and Parker, 1998). In migrating Walker 256 carcinosarcoma cells, uropods are negatively regulated by PKC (Niggli et al, 1996;Rossy et al, 2007). We therefore examined whether the localisation of the ezrin spot was affected by the DAG analogue phorbol 12- Intensity plots of these images are shown in G. (H)Cells expressing ezrin-GFP were stained with TexasRed-phalloidin (TR-phalloidin) to visualise F-actin.…”

mentioning

confidence: 99%

See 1 more Smart Citation

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

Lorentzen

Bamber

Sadok

et al. 2011

Journal of Cell Science

108

111

View full text Add to dashboard Cite

SummaryMelanoma cells can switch between an elongated mesenchymal-type and a rounded amoeboid-type migration mode. The rounded 'amoeboid' form of cell movement is driven by actomyosin contractility resulting in membrane blebbing. Unlike elongated A375 melanoma cells, rounded A375 cells do not display any obvious morphological front-back polarisation, although polarisation is thought to be a prerequisite for cell movement. We show that blebbing A375 cells are polarised, with ezrin (a linker between the plasma membrane and actin cytoskeleton), F-actin, myosin light chain, plasma membrane, phosphatidylinositol (4,5)-bisphosphate and b1-integrin accumulating at the cell rear in a uropod-like structure. This structure does not have the typical protruding shape of classical leukocyte uropods, but, as for those structures, it is regulated by protein kinase C. We show that the ezrin-rich uropod-like structure (ERULS) is an inherent feature of polarised A375 cells and not a consequence of cell migration, and is necessary for cell invasion. Furthermore, we demonstrate that membrane blebbing is reduced at this site, leading to a model in which the rigid ezrin-containing structure determines the direction of a moving cell through localised inhibition of membrane blebbing. Journal of Cell ScienceIn confined spaces, blebbing cells can move forward without adhesion by a process termed 'chimneying' (Hawkins et al., 2009;Malawista and de Boisfleury Chevance, 1997). In fibrillar threedimensional matrices, such as collagen gels, blebs can expand into pores in the matrix, causing the squeezing phenotype observed in amoeboid lymphocytes and cancer cells in collagen gels (Haston and Shields, 1984;Wolf et al., 2003a). Cells that form small blebs can use them to 'elbow' their way through the meshwork of the ECM (Friedl and Wolf, 2009). The site of bleb formation determines the direction of cell movement (Keller and Bebie, 1996). However, some cells, such as A375 melanoma cells, form multiple blebs over the membrane and would therefore not be able to move directionally. However, A375 melanoma cells are able to invade into a collagen matrix and move in vitro and in vivo (Sahai and Marshall, 2003;Sanz-Moreno et al., 2008). As asymmetry is a prerequisite of cell movement (Petrie et al., 2009), this suggests some form of cellular polarisation. To understand cell movement driven by multiple blebs, we investigated polarisation in blebbing 'amoeboid' A375 melanoma cells. We show that A375 cells form an ezrin-rich uropod-like structure (which we call the ERULS) at their cell rear, and that this is required for invasion into a three-dimensional matrix. We present a model whereby accumulation of ezrin, membrane and actin in this structure reflects strong connections between the plasma membrane and the submembrane cortex, leading to the formation of a rigid structure that inhibits blebbing at the back of the cell. This leads to net movement of the cell in the direction opposite to the uropod-like structure. Our model explains how cells can move ...

show abstract

Section: Rock and Novel Pkc Isoforms Regulate The Erulsmentioning

confidence: 99%

mentioning

confidence: 99%

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

Lorentzen

Bamber

Sadok

et al. 2011

Journal of Cell Science

108

111

View full text Add to dashboard Cite

show abstract

“…Cytosolic and membrane fractions were then isolated as described previously. 24 Fractions were solubilized in 60 l sample buffer containing 1% SDS, 50 mM DTT, 15% glycerol, 62.5 mM Tris/ HCl, pH 6.8, and 0.001% bromphenol blue by incubation at 95°C for 5 min. Fractions were electrophoresed through a 5% to 10% SDS-polyacrylamide gradient gel followed by transblotting to nitrocellulose using a Genie blotter (Idea Scientific, Minneapolis, MN).…”

Section: Adp Ribosylationmentioning

confidence: 99%

The Rho/Rho-kinase and the phosphatidylinositol 3-kinase pathways are essential for spontaneous locomotion of Walker 256 carcinosarcoma cells

Wicki

Niggli

2001

Int. J. Cancer

Self Cite

View full text Add to dashboard Cite

“…Several studies document that activation of PKC is a stop signal for tumor cell locomotion, but other studies document that PKC activation can stimulate invasion. Data supporting the view that activation of PKC is a stop signal for tumor cell locomotion and that decreased PKC levels stimulate locomotion have been obtained in Walker carcinosarcoma cells (Keller et al, 1985, Keller et al, 1989; Niggli et al, 1996; Wicki and Niggli, 1999), human keratinocytes (Ando et al, 1993) and rat glioma cells (Janik et al, 1992). Similar findings have been obtained in human neutrophils or lymphocytes (Roos et al, 1987; Trachsel and Keller, 1995).…”

mentioning

confidence: 79%

Protein kinase C isoforms involved in regulation of cell shape and locomotion of human fibrosarcoma HT1080 cells

Keller

Hunziker

Sordat³

et al. 2000

Int. J. Cancer

View full text Add to dashboard Cite

The role of protein kinase C (PKC) isoforms in the regulation of cell shape [switch between fibroblast‐like and crescent shape (CS)] and of locomotion of human fibrosarcoma HT1080 cells has been investigated. The PKC activator phorbol myristate acetate (PMA) induced the transition of elongated fibroblast‐like cells into CS cells and stimulated locomotion. Both responses to PMA were inhibited by the PKC inhibitor Ro 31‐8220. Analysis of the time course showed that stimulation of shape changes (formation of CS cells) and locomotor activity (increase in the proportion and speed of locomoting cells) was maximal in the early phase of the response (up to 2.5 hr) and significantly decreased later (15 to 21 hr). CS formation and stimulated locomotion correlated closely with a marked redistribution from the cytosol to the membrane of PKC isoforms α, β1 and γ in the early phase (0.5 to 2 hr) following activation with PMA. The subsequent reduction of the proportion of CS cells and of cell locomotion correlated with down‐regulation of these isoforms in the second phase (16 to 21 hr). In contrast, the total amount and distribution of PKC β2 remained almost unchanged with 10–8 M PMA up to 21 hr. Furthermore, changes in shape and locomotion did not correlate with the responses of PKC δ to PMA. Inhibition of PMA‐stimulated locomotion by the more specific inhibitor Gö 6976 is consistent with a role of PKC α and β1 in this response. Ro 31‐8220 alone induced a moderate down‐regulation of PKC isoforms α and δ, but it also inhibited the more pronounced down‐regulation of these isoforms by PMA. Our results indicate that activation of PKC isoforms α, γ and β1, but not β2 or δ, stimulates locomotion and formation of CS cells in human fibrosarcoma HT1080 cells. Int. J. Cancer 88:195–203, 2000. © 2000 Wiley‐Liss, Inc.

show abstract

Inhibition of protein kinase C-dependent protein phosphorylation correlates with increased polarity and locomotion in Walker 256 carcinosarcoma cells

Abstract: P~r m o t t t m l n t e m a t l o n a t U n a n~n s i C a n c e r%

Cited by 11 publications

References 16 publications

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

An ezrin-rich, rigid uropod-like structure directs movement of amoeboid blebbing cells

The Rho/Rho-kinase and the phosphatidylinositol 3-kinase pathways are essential for spontaneous locomotion of Walker 256 carcinosarcoma cells

Protein kinase C isoforms involved in regulation of cell shape and locomotion of human fibrosarcoma HT1080 cells

Contact Info

Product

Resources

About